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Plasticity and cond...
Plasticity and conditional essentiality of modification enzymes for domain V of Escherichia coli 23S ribosomal RNA
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- Liljeruhm, Josefine (author)
- Uppsala universitet,Institutionen för cell- och molekylärbiologi
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- Leppik, Margus (author)
- Univ Tartu, Dept Mol Biol, EE-51010 Tartu, Estonia.
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- Bao, Letian (author)
- Uppsala universitet,Molekylärbiologi
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- Truu, Triin (author)
- Univ Tartu, Dept Mol Biol, EE-51010 Tartu, Estonia.
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- Calvo-Noriega, Maria (author)
- Uppsala universitet,Institutionen för cell- och molekylärbiologi
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- Freyer, Nicola S. (author)
- Uppsala Univ, Dept Cell & Mol Biol, S-75124 Uppsala, Sweden.
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- Liiv, Aivar (author)
- Univ Tartu, Dept Mol Biol, EE-51010 Tartu, Estonia.
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- Wang, Jinfan (author)
- Uppsala Univ, Dept Cell & Mol Biol, S-75124 Uppsala, Sweden.
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- Blanco, Ruben Crespo (author)
- Uppsala Univ, Dept Cell & Mol Biol, S-75124 Uppsala, Sweden.
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- Ero, Rya (author)
- Univ Tartu, Dept Mol Biol, EE-51010 Tartu, Estonia.
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- Remme, Jaanus (author)
- Univ Tartu, Dept Mol Biol, EE-51010 Tartu, Estonia.
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- Forster, Anthony C. (author)
- Uppsala universitet,Molekylärbiologi,Uppsala Univ, Dept Cell & Mol Biol, S-75124 Uppsala, Sweden.
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(creator_code:org_t)
- 2022-03-08
- 2022
- English.
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In: RNA. - : Cold Spring Harbor Laboratory Press (CSHL). - 1355-8382 .- 1469-9001. ; 28:6, s. 796-807
- Related links:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
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- Escherichia coli rRNAs are post-transcriptionally modified at 36 positions but their modification enzymes are dispensable individually for growth, bringing into question their significance. However, a major growth defect was reported for deletion of the RlmE enzyme, which abolished a 2 ' O methylation near the peptidyl transferase center (PTC) of the 23S rRNA. Additionally, an adjacent 80-nt "critical region" around the PTC had to be modified to yield significant peptidyl transferase activity in vitro. Surprisingly, we discovered that an absence of just two rRNA modification enzymes is conditionally lethal (at 20 degrees C): RlmE and RluC. At a permissive temperature (37 degrees C), this double knockout was shown to abolish four modifications and be defective in ribosome assembly, though not more so than the RlmE single knockout. However, the double knockout exhibited an even lower rate of tripeptide synthesis than did the single knockout, suggesting an even more defective ribosomal translocation. A combination knockout of the five critical-region-modifying enzymes RluC, RlmKL, RlmN, RlmM, and RluE (not RlmE), which synthesize five of the seven critical-region modifications and 14 rRNA and tRNA modifications altogether, was viable (minor growth defect at 37 degrees C, major at 20 degrees C). This was surprising based on prior in vitro studies. This five-knockout combination had minimal effects on ribosome assembly and frameshifting at 37 degrees C, but greater effects on ribosome assembly and in vitro peptidyl transferase activity at cooler temperatures. These results establish the conditional essentiality of bacterial rRNA modification enzymes and also reveal unexpected plasticity of modification of the PTC region in vivo.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Keyword
- E
- coli
- modification enzymes
- rRNA
- tRNA
- translation
Publication and Content Type
- ref (subject category)
- art (subject category)
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RNA
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Liljeruhm, Josef ...
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Leppik, Margus
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Bao, Letian
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Truu, Triin
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Calvo-Noriega, M ...
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Freyer, Nicola S ...
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show more...
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Liiv, Aivar
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Wang, Jinfan
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Blanco, Ruben Cr ...
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Ero, Rya
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Remme, Jaanus
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Forster, Anthony ...
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- NATURAL SCIENCES
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RNA
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Uppsala University